Numerical count of membrane proteins

Hello everyone,
I’m doing some exercise to better understand imageJ.
My goal is to count the number of staining membrane.
I took this image from internet to do exercise.
Could someone explain me which procedure he would use if he were in my situation?
I know i could easily do it with CellProfiler but this is not my goal.
Thanks to everyone who will want to help me.

CellBrite-Blue-fixed-cells

I wasn’t quite sure what you are asking for, based on your description.
Are you trying to count the number of cells positive for a membrane stain? If so, probably some sort of cleanup (standard deviation filter?)-> low pass threshold to find the black areas between membranes-> Analyze particles with some sort of size threshold. You may also want to take each ROI, after the Analyze particles step, and expand it several pixels to see if that area contains “bright enough” pixels to be considered membrane staining.

Or are you quantifying the relative amounts of protein between two images (which would, at the least, require some sort of negative control and background removal/thresholding)?

I wanted to count the number of cells positive for a membrane stain. I also used the strategy you suggested me. I wanted to make sure that the method I used was the best possible. thanks for your time :slightly_smiling_face: